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Inukai Tsuyoshi

Research Faculty of Agriculture　Fundamental AgriScience Research　Agrobiology and BioresourcesLecturer

Researcher basic information

■ Degree
  • 博士(農学）, 北海道大学
■ URL
researchmap URLホームページURL■ Various IDs
J-Global ID■ Research Keywords and Fields
Research Keyword
  • Brassica rapa
  • イネいもち病
  • Turnip mosaic virus
  • デンプン合成
  • 圃場抵抗性
  • 病徴制御
  • 登熟
  • Oryza Sativa
Research Field
  • Environmental Science/Agriculture Science, Science in plant genetics and breeding
■ Educational Organization

Research activity information

■ Awards
  • Apr. 2017, 日本植物病理学会, JGPP Outstanding Paper Award
    Identification of the TuNI gene causing systemic necrosisin Arabidopsis ecotype Ler infected with Turnip mosaic virus and characterization of its expression（JGPP, 2015, 81:180–191)
    Jinyan Liu •;Bo Min Kim;Yo-hei Kaneko;Tsuyoshi Inukai;Chikara Masuta
■ Papers
■ Other Activities and Achievements
■ Syllabus
  • 農業生物学特論, 2024年, 修士課程, 農学院
  • 農業生物学特論演習, 2024年, 修士課程, 農学院
  • 一般教育演習(ﾌﾚｯｼｭﾏﾝｾﾐﾅｰ), 2024年, 学士課程, 全学教育
  • 植物感染生理学, 2024年, 学士課程, 農学部
■ Affiliated academic society
  • PHYTOPATHOLOGICAL SOCIETY OF JAPAN
  • JAPANESE SOCIETY OF BREEDING
■ Research Themes
  • イネ感染特異的タンパク質PR1bのアポプラストにおける挙動並びに防御機能の解明
    科学研究費助成事業
    01 Apr. 2023 - 31 Mar. 2026
    犬飼 剛
    コシヒカリに見出されたいもち病高度罹病性QTLの責任遺伝子が機能喪失したPR1bであることを確かめるため、トウモロコシのユビキチンプロモーターに日本晴由来の野生型PR1bをつないだコンストラクトを作成し、これをpBract発現ベクターに組み込んでアグロバクテリウム法によりコシヒカリに導入した。形質転換体の世代を進め、本年度T2固定系統が得られたことから、これらを用いて噴霧接種法によるいもち病抵抗性の検定を行った。その結果、T2固定系統において抵抗性病斑の割合の増加など、量的な抵抗性の増大が認められた。この結果から、コシヒカリがもついもち病高度罹病性QTLの責任遺伝子は機能喪失したPR1bである可能性が高いと考えられた。また、抵抗性病斑の割合が増加する傾向が認められたことから、PR1bはいもち病菌の増殖を阻害するか、あるいは植物自身の抵抗性を増大させる機能があることが示唆された。一方、日本晴のPR1bに関するコシヒカリの準同質遺伝子系統と比較した場合、抵抗性の程度に大きな違いは認められず、過剰発現により効果が高まることはないと考えられた。
    PR1タンパクのC末端11アミノ酸は、プロテアーゼによって切り出されCAPEと呼ばれるペプチドシグナルとして防御反応の誘導に働くことが知られているが、イネのPR1bにもCAPEと相同な配列が存在する。イネPR1b由来のCAPEの機能を調べるため、人工合成したCAPEをイネ葉に施与していもち病菌に対する抵抗性が誘導されるか接種試験を行って調べたところ、処理葉において明瞭な抵抗性の増大は認められなかった。
    日本学術振興会, 基盤研究(C), 北海道大学, 23K05160
  • イネいもち病圃場抵抗性遺伝子Pid3-I1の作用機作の解明
    Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Apr. 2020 - Mar. 2023
    犬飼 剛
    Pid3-I1はいもち病菌に対して量的に働く抵抗性遺伝子である。このPid3-I1の効果をそれぞれCO39とコシヒカリの遺伝的背景下で比較すると、CO39とコシヒカリの罹病程度は同定程度であるにも関わらず、コシヒカリの遺伝的背景下ではPid3-I1の効果がCO39におけるそれと比べて低い傾向にあった。これはPid3-I1と相互作用して抵抗性を増大させる因子があることを示唆している。この因子を同定する目的で、いもち病菌感染時における防御関連遺伝子の発現パターンをCO39とコシヒカリで比較したところ、コシヒカリではPR1bの発現が全く誘導されないことを見いだした。PR1タンパク質は抗菌タンパク質として病原菌類に作用するだけでなく、タンパク質C末端の11アミノ酸がプロテアーゼによって切断され、CAPE1ペプチドエリシターとして抵抗性の誘導に関わることが他種植物で報告されており、イネにおいてPid3-I1を介した抵抗性誘導経路がCAPE1を介した経路とクロストークしている可能性が示唆された。本研究では、まずコシヒカリにおいてPR1bが発現しない原因を調べた。その結果、PR1bのアミノ酸コード領域にレトロトランスポゾン様因子が挿入されており、この挿入変異によって遺伝子の機能が喪失していることが明らかになった。また、このPR1b座における多型をマーカー化して日本の栽培品種における拡がりを調べたところ、主要品種のほとんどがこの変異をもつことが示された。このため、現在の日本品種にPid3-I1を導入しても十分な圃場抵抗性の向上が期待できない可能性がある。今後、PR1bをコシヒカリに導入した過剰発現体を作成し、イネにおけるPR1bの機能を解析するとともにPid3-I1との相互作用を解析する。
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 20K05968
  • イネいもち病圃場抵抗性遺伝子qBRM6.2の単離と機能解析
    科学研究費補助金(基盤研究(C))
    2016 - 2018
    犬飼剛
    文部科学省, Principal investigator, Competitive research funding
  • Analysis of induction mechanism for ascorbic acid accumulation as antiviral reaction
    Grants-in-Aid for Scientific Research
    2012 - 2014
    INUKAI Tsuyoshi; MASUTA Chikara; FUJIWARA Ayaka
    In this study, we first showed that endogenous ascorbic acid (AsA) functioned as an antiviral chemical to TuMV in Brassica rapa and Arabidopsis thaliana. Next, it was shown that the AsA accumulation was induced by TuMV infection in the B. rapa resistant cultivar Aki-masari. This AsA accumulation was due to suppression of AsA oxidation by ascorbate peroxidase (APX) and ascorbate oxidase and activation of AsA recycling by dehydroascorbate reductase (DHAR). Because methyl jasmonate (MeJA) suppressed the APX activity and enhanced the DHAR activity; JA derivatives including JA-isoleucine, tuberonic acid and tuberonic acid glucoside were increased together with AsA, the induction of the AsA accumulation in Aki-masari appeared to be mediated by the JA-dependent signaling pathway suggesting that the AsA accumulation was induced as a defense reaction against virus.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, Principal investigator, Competitive research funding, 24580001
  • Development of symptom control system for plant virus disease -Brassica crops and Turnip mosaic virus-
    Grants-in-Aid for Scientific Research(基盤研究(C))
    2008 - 2010
    Tsuyoshi INUAKAI; Chikara MASUTA; Ayaka FUJIWARA
    In the pathosystem of Brassica rapa and Turnip mosaic virus (TuMV), the type of symptoms expressed by susceptible plants are determined by the gene combinations between the host cultivar and virus strain. In this study, we found that the resistant reaction and symptoms such as systemic lethal necrosis, leaf malformation and mosaic were differentially determined, depending on the combinations of the genotypes for the Rnt1 locus and the viral CI gene.
    Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(C), 北海道大学, Principal investigator, Competitive research funding, 20580002
  • Mechanism for induction of symptom by virus infection in plant
    Grants-in-Aid for Scientific Research(基盤研究(C))
    2005 - 2006
    Tsuyoshi INUKAI
    In the pathosystems of Turnip mosaic virus (TuMV) and Brassica crops, various symptoms are developed such as mosaic and necrosis leading to loss of yield and product quality. Although necrotic symptom causes more severe damage in plants infected by TuMV, the induction mechanism of necrosis is poorly understood so far. We previously identified the symptom determinant TuNI in Arabidopsis thaliana, a model species of the family Brassicaceae. The TuNI gene directly or indirectly interacts to a TuMV factor and induces veinal necrosis in Arabidopsis. In this study it was shown that the necrotic s...
    Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(C), 北海道大学, Principal investigator, Competitive research funding, 17580001
  • 栽培イネにおける登熟生理に関する育種学的研究
    科学研究費補助金(若手研究(B))
    2002 - 2003
    犬飼 剛
    本年度はインディカ型及びジャポニカ型のイネ品種間で認められる登熟速度の差がどのような要因によって生じているのか明らかにする目的で研究を行った。登熟速度の異なるインディカ型及びジャポニカ型のイネ品種間で胚乳におけるデンプンの蓄積速度及びデンプン合成関連酵素の活性を比較したところ、インディカ型品種はジャポニカ型品種に比べて登熟初期における可溶性デンプン合成酵素の活性が2倍程度高く、その結果デンプンの蓄積速度もインディカ型品種の方で大きくなることを明らかにした。デンプンの組成別に見ると、登熟初期ではアミロースの蓄積速度に品種間で大きな差はないのに対し、アミロペクチンの蓄積速度はインディカ型品種の方で大きかった。このことから登熟初期におけるデンプン蓄積速度の品種間差は主にアミロペクチンの合成速度の差に起因すると考えられた。また、デンプンに占めるアミロースの比率で見ると、登熟初期ではインディカ型品種とジャポニカ型品種で差がないのに対し登熟後期では品種間で10%近く差を生じており、コメの品質に大きな影響を与えるアミロースに関してその含量だけではなく蓄積パターンにもインディカ型品種とジャポニカ型品種とでは大きな違いがあることが明らかとなった。以上の結果から、インディカ型品種に見られる早熟性の大きな要因としてアミロペクチン合成過程における合成速度の変異が考えられ、またこの変異はデンプンの組成...
    文部科学省, 若手研究(B), 北海道大学, Principal investigator, Competitive research funding, 14760001
  • Study on the induction system of host specificity mutations in Magnaporthe grisea,: avirulence genes and recombinational repair genes
    Grants-in-Aid for Scientific Research(基盤研究(A))
    1999 - 2001
    Fusao TOMITA; 犬飼 剛; 中島 敏彦; 石井 千津; 安田 伸子; 曾根 輝雄; 辻本 雅子
    (1) Analysis of avirulence genes in M. griseaThree avirulence genes, Avr-pii, Avr-pia, Avr-Hattan3 was investigated using a cross between two Yunnang isolates. These avirulence genes were mapped onto the genetic map of the fungus. Along the Avr-Hattan 3, a BAC contig of 600kb in length was constructed. On the other hand, Avr-pia was identified also from a cross between Japanese isolate and rare hermaphloditic isolate Guy11. By utilization of the host specificity mutant, a DNA fragment named PM-01, which was tightly linked to the Avr-pia locus was isolated. One of the cosmid clone containing...
    Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(A), 北海道大学, Coinvestigator not use grants, Competitive research funding, 11306007
  • The use of molecular information for plant breeding
    Grants-in-Aid for Scientific Research(基盤研究(B))
    1999 - 2000
    Yoshio SANO; 三上 哲夫; 犬飼 剛
    The objectives of the present study is to select and create new genes by means of molecular approaches in rice. Firstly, the experiments were carried out regarding the diversification of cis-acting elements of the Wx gene whose expressions have a role for eating quality. By comparing DNA sequences and recombination frequency, it was found that the region in the Wx gone is recombinationally active and it is possible to recombine different cis-acting elements which are present among rice cultivars. Secondly, the experiments were carried out to induce trans-acting mutant genes. The Wx gene exp...
    Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(B), 北海道大学, Coinvestigator not use grants, Competitive research funding, 11556001
  • Studies on the differentiation of genepools in rice
    Grants-in-Aid for Scientific Research(基盤研究(A))
    1997 - 2000
    Yoshio SANO; 金澤 章; 犬飼 剛; 貴島 祐治
    Plant breeding is mainly to select favorable gene combinations from naturally occurring variants through hybridization. Since species have the primary genepool which is used as the genetic materials for their adaptation, the differential genepool is closely related with the difficulties in hybridization breeding. The objectives of the present study is to reveal the differentiated genepool among rice taxa and to examine the causal factors. The genic diversity was examined regarding to Wx and C genes controlling amylose content and anthocyanin coloration since these variations are distinct am...
    Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(A), 北海道大学, Coinvestigator not use grants, Competitive research funding, 09306001
  • イネゲノム中に見出されたMAR様ミニサテライトの構造・機能解析
    ホクト生物科学振興財団奨励金
    2000
    犬飼剛
    Principal investigator, Competitive research funding
  • Fine structural analysis of rice wx and the application for breeding
    Grants-in-Aid for Scientific Research(基盤研究(C))
    1998 - 1999
    Tsuyoshi INUKAI; 佐野 芳雄
    In plant genomes as well as other eukaryotic genomes meiotic recombination does not occur uniformly. At the level of the gene, high recombination frequencies are often observed within genetic loci in maize, but this feature of intragenic recombination is not seen at the csr1 locus in Arabidopsis. These observations suggest that meiotic recombination in plant genomes varies considerably among species. In the present study we investigated meiotie recombination at the wx locus in rice. The mutation sites of wx mutants induced by ethyl methanesulfonate (EMS) treatment or gamma-ray irradiation a...
    Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(C), 北海道大学, Principal investigator, Competitive research funding, 10660001
  • イネのwx遺伝子座における温度非感受性変異体の作出
    タカノ農芸化学研究助成財団研究助成金
    1997
    犬飼剛
    Principal investigator, Competitive research funding
  • 普通野生稲集団におけるいもち病抵抗性に関する遺伝的構造の解明
    稲盛財団助成金
    1997
    犬飼剛
    Principal investigator, Competitive research funding
  • Molecular Mechanism of Cytoplasmic Male Sterility in Sugarbeet (Beta vulgaris L.)
    Grants-in-Aid for Scientific Research
    1994 - 1995
    MIKAMI Tetsuo; INUKAI Tuyoshi; KUBO Tomohiko
    To study the molecular basis of cytoplasmic male sterility (CMS) in sugarbeet, we have conducted a detailed comparison of the mitochondrial genomes of two sugarbeet lines, one containing a fertile (N) cytoplasm and the other a male sterile one. The two genomes have approximately 330kb of common sequences but there is considerable variation in sequence organization which can be accounted for by genome rearrangements.
    Transcripts for known mitochondrial genes and regions of the two sugarbeet genomes known to be rearranged relative to one another were compared among male-fertile, CMS and nuclear-restored sugarbeet genotypes. In most instances, identical transcript patterns were observed for all three, mtRNAs (prepared from leaves or flower buds), indicating that rearrangement events did not occur within transcriptional units. Altered transcript patterns were, however, identified for six genes, coxI,coxII,atpA,rps3, orf324, and orf1.
    Interestingly, the coxI transcripts were found to be significantly affected by nuclear background, whereas unclear background has no effect on the transcripts of other five genes.
    The results suggest that nuclear-restorer gene (s) have an effect on coxI transcript patterns and implicate coxI in sugarbeet male sterility.
    Japan Society for the Promotion of Science, Grant-in-Aid for General Scientific Research (B), HOKKAIDO UNIVERSITY, 06454039
  • Re-evaluation of alien wild genes in rice
    Grants-in-Aid for Scientific Research
    1993 - 1995
    SANO Yoshio; INUKAI Tsuyoshi; TAKAMURE Itsurou
    Considering that genetic erosion is occurring at an accelerating rate through the world, it is urgent to evaluate and use alien genes. To overcome difficulties associated with gene transfer from alien species, we must learn more about the basic knowledge on crop genepools. Here, we examined constraints in using wild rice relatives in breeding.
    Unidirectional cross-incompatibility was detected when a segment of chromosome 6 was introduced from wild rice into a cultivar. When pollinated by Japonica type, the plant with the segment showed reduced seed setting while the reciprocal showed normal seed setting. Cytohistological observations revealed that retardation of the endosperm caused abortion of seeds. Genetic experiments revealed that a gene controlling incompatibility in the female parent as well as a gene (s) controlling compatibility in the male parent was located on the segment. Furthermore, Japonica type tended to show reduced seed setting compared to Indica type and wild rice. A segment of chromosome 7 was also examined after introducing it into a cultivated form from a common wild rice. The genetic diver-gence on the segment is needed to be studied since weedy forms frequently show red pericarps even after introgression, suggesting the adaptive significances of the segment under natural conditions. Two dominant genes were detacted to be located on the segment. One was responsible for short stature only in the vegetative phase. The other gene was responsible for photosensitivity. Distorted segreations were detected and the introduced segment seemed to be quickly eliminated as the plants become similar to cultivated type. Thus, internal constraints between different taxa are needed to be studied for efficient uses of alien genes.
    Japan Society for the Promotion of Science, Grant-in-Aid for General Scientific Research (B), 05454041
  • イネにおけるいもち病安定抵抗性の遺伝機構の解明
    科学研究費補助金(奨励研究(A))
    1994 - 1994
    犬飼 剛
    イネいもち病に対して安定した抵抗性を示すことで知られている西アフリカのジャポニカ品種Moroberekanのいもち病抵抗性の遺伝機構を解明するため、Moroberekan中に複数存在する抵抗性遺伝子に関する準同質遺伝子系統を作成し、抵抗性遺伝子の同定を行うことを目的に実験を行っている。現在までの実施結果は以下の通りである。高度羅病性インディカ品種CO39とMoroberekanとの組合せより育成された組換近交系の中から、Moroberekan由来の真性抵抗性遺伝子もしくは圃場抵抗性遺伝子をもつと推定され、なおかつ遺伝的背景がCO39に近似した系統を選抜した。次に選抜した系統中の抵抗性遺伝子の遺伝分析を行うため、CO39との交配を行いF_2-F_3を養成した。F_2世代では、後の選抜のために出穂期、稈長等の調査を行うとともに、目的とする抵抗性遺伝子に連鎖すると推定されるRFLPマーカーの分離を調べた。現在F_3世代で抵抗性遺伝子の分離を調査中である。これらの結果より、複数存在する抵抗性遺伝子のいずれを選抜系統がもつか明らかにするとともに、遺伝子分析用に養成したF_3系統の中から、形態選抜及びmarker-assisted selectionにより、各抵抗性遺伝子に関する準同質遺伝子系統を選抜する。これらの準同質遺伝子系統のうち、真性抵抗性遺伝子に関するものについてはいもち病菌レ...
    文部科学省, 奨励研究(A), 北海道大学, Principal investigator, Competitive research funding, 06760002
  • Regulation of fertilization and pollen selection in rice
    Grants-in-Aid for Scientific Research(一般研究(C))
    1992 - 1993
    Masashiko MAEKAWA; 犬飼 剛
    The results obtained in the sutdy are as follows ;(1) Function of gametophyte geneFunction of gametophyte gene, g^ was inverstigated by using the isogeneic line, T-65 g^ which possesses g^ in T-65.In the same-timing pollination of T-65 and T-65 g^, pollens with g^ show fertilization rate of about 1/3 compared with normal pollens. In the time-lagged pollination of pollens with g^, lately pollinated pollens with g^ show fertilization rate of below 1/4 compared with formerly pollinated normal pollens. This result suggests that thers is differential elongation...
    Ministry of Education, Culture, Sports, Science and Technology, 一般研究(C), 北海道大学, Coinvestigator not use grants, Competitive research funding, 04660001